LOCUS       sequence-176830-        4999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4999)
  AUTHORS   Zhou BP, Deng J, Xia W, Xu J, Li YM, Gunduz M, Hung MC
  TITLE     Dual regulation of Snail by GSK-3beta-mediated phosphorylation in 
            control of epithelial-mesenchymal transition.
  JOURNAL   Nat Cell Biol. 2004 Oct . 6(10):931-40.
  PUBMED    15448698
REFERENCE   2  (bases 1 to 4999)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Cell 
            Biol. 2004 Oct . 6(10):931-40."
COMMENT     Sequence Label: Flag Snail 6SA
FEATURES             Location/Qualifiers
     source          1..4999
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     64..83
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        197..301
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(326..346)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        370..580
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        392..588
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      439..574
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             623..1417
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             623..1417
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(677..696)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1287..1306
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(1605..1624)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1649..1696
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      2025..2613
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2514..2533
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2788..3091
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3092..3295
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3246..3266
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        3341..3359
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     regulatory      3394..3403
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3403..3426
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(4293..4312)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4294..4312)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    4586..4707
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4623..4642)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4677..4696
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(join(4714..4999,1..170))
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4851..4872)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
       61 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
      121 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt caggtggcac
      181 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
      241 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagaa
      301 tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
      361 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
      421 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
      481 catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc ctctgagcta
      541 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaga tcgatcaaga
      601 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
      661 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
      721 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
      781 gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac
      841 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
      901 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
      961 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     1021 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     1081 cgatcaggat gatctggacg aagaacatca ggggctcgcg ccagccgaac tgttcgccag
     1141 gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     1201 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     1261 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagaacttgg
     1321 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     1381 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     1441 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     1501 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     1561 gatctcatgc tggagttctt cgcccaccct agggggaggc taactgaaac acggaaggag
     1621 acaataccgg aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggt
     1681 gttgggtcgt ttgttcataa acgcggggtt cggtcccagg gctggcactc tgtcgatacc
     1741 ccaccgagac cccattgggg ccaatacgcc cgcgtttctt ccttttcccc accccacccc
     1801 ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg gggcggcagg ccctgccata
     1861 gcctcaggtt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     1921 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     1981 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     2041 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     2101 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     2161 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     2221 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     2281 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     2341 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     2401 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     2461 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     2521 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     2581 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     2641 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
     2701 gtggataacc gtattaccgc catgcattag ttattaatag taatcaatta cggggtcatt
     2761 agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg gcccgcctgg
     2821 ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc ccatagtaac
     2881 gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt
     2941 ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca atgacggtaa
     3001 atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta cttggcagta
     3061 catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt acatcaatgg
     3121 gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg
     3181 gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc
     3241 attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt
     3301 agtgaaccgt cagatccgct agcgattacg ccaagctcga aattaaccct cactaaaggg
     3361 aacaaaagct ggagctccac cgcggtggcg gccgccacca tggattacaa ggatgacgac
     3421 gataagagcc cgggcggatc cccgcgctct ttcctcgtca ggaagccctc cgaccccaat
     3481 cggaagccta actacagcga gctgcaggac tctaatccag agtttacctt ccagcagccc
     3541 tacgaccagg cccacctgct ggcagccatc ccacctccgg agatcctcaa ccccaccgcc
     3601 tcgctgccaa tgctcatctg ggactctgtc ctggcgcccc aagcccagcc aattgcctgg
     3661 gcctcccttc ggctccagga gagtcccagg gtggcagagc tgacctccct gtcagatgag
     3721 gacgctggga aaggcgccca gccccccagc ccacccgcac cggctcctgc gtccttctcc
     3781 gcgacgtcag tcgcttcctt ggaggccgag gcctatgctg ccttcccagg cttgggccaa
     3841 gtgcccaagc agctggccca gctctctgag gccaaggatc tccaggctcg aaaggccttc
     3901 aactgcaaat actgcaacaa ggaatacctc agcctgggtg ccctcaagat gcacatccga
     3961 agccacacgc tgccctgcgt ctgcggaacc tgcgggaagg ccttctctag gccctggctg
     4021 ctacaaggcc atgtccggac ccacactggc gagaagccct tctcctgtcc ccactgcagc
     4081 cgtgccttcg ctgaccgctc caacctgcgg gcccacctcc agacccactc agatgtcaag
     4141 aagtaccagt gccaggcgtg tgctcggacc ttctcccgaa tgtccctgct ccacaagcac
     4201 caagagtccg gctgctcagg atgtccccgc tgactcgagg gggggcccgg taccttaatt
     4261 aattaaggta ccaggtaagt gtacccaatt cgccctatag tgagtcgtat tacaattcac
     4321 tcgatcgccc ttcccaacag ttgcgcagcc tgaatggcga atggagatcc aatttttaag
     4381 tgtataatgt gttaaactac tgattctaat tgtttgtgta ttttagattc acagtcccaa
     4441 ggctcatttc aggcccctca gtcctcacag tctgttcatg atcataatca gccataccac
     4501 atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga acctgaaaca
     4561 taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg gttacaaata
     4621 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     4681 tttgtccaaa ctcatcaatg tatcttaacg cgtaaattgt aagcgttaat attttgttaa
     4741 aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc gaaatcggca
     4801 aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt ccagtttgga
     4861 acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa accgtctatc
     4921 agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc
     4981 gtaaagcact aaatcggaa
//
